• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of cytotechspringer.comThis journalToc AlertsSubmit OnlineOpen Choice
Cytotechnology. Jul 1999; 30(1-3): 211–226.
PMCID: PMC3449946

Embryonic stem cell differentiation models: cardiogenesis, myogenesis, neurogenesis, epithelial and vascular smooth muscle cell differentiation in vitro


Embryonic stem cells, totipotent cells of the early mouse embryo, were established as permanent cell lines of undifferentiated cells. ES cells provide an important cellular system in developmental biology for the manipulation of preselected genes in mice by using the gene targeting technology. Embryonic stem cells, when cultivated as embryo-like aggregates, so-called ‘embryoid bodies’, are able to differentiate in vitro into derivatives of all three primary germ layers, the endoderm, ectoderm and mesoderm. We established differentiation protocols for the in vitro development of undifferentiated embryonic stem cells into differentiated cardiomyocytes, skeletal muscle, neuronal, epithelial and vascular smooth muscle cells. During differentiation, tissue-specific genes, proteins, ion channels, receptors and action potentials were expressed in a developmentally controlled pattern. This pattern closely recapitulates the developmental pattern during embryogenesis in the living organism. In vitro, the controlled developmental pattern was found to be influenced by differentiation and growth factor molecules or by xenobiotics. Furthermore, the differentiation system has been used for genetic analyses by ‘gain of function’ and ‘loss of function’ approaches in vitro.

Keywords: cardiogenesis, cell differentiation, gene expression, mouse embryonic stem cells, myogenesis, neurogenesis

Full Text

The Full Text of this article is available as a PDF (805K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bagutti C, Wobus AM, Fässler R, Watt FM. Differentiation of embryonal stem cells into keratinocytes: Comparison of wild-type and β1 integrin-deficient cells. Dev Biol. 1996;179:184–196. doi: 10.1006/dbio.1996.0250. [PubMed] [Cross Ref]
  • Bain G, Kitchens D, Yao M, Huettner JE, Gottlieb DI. Embryonic stem cells express neuronal properties in vitro. Dev Biol. 1995;168:342–357. doi: 10.1006/dbio.1995.1085. [PubMed] [Cross Ref]
  • Blank RS, Swartz EA, Thompson MM, Olson EN, Owens GK. A retinoic acid-induced clonal cell line derived from multipotential P19 embryonal carcinoma cells expresses smooth muscle characteristics. Circ Res. 1995;76:742–749. [PubMed]
  • Boudjelal M, Taneja R, Matsubara S, Bouillet P, Dolle P, Chambon P. Overexpression of Stra13, a novel retinoic acid-inducible gene of the basic helix-loop-helix family, inhibits mesodermal and promotes neuronal differentiation of P19 cells. Genes Dev. 1997;11:2052–2065. [PMC free article] [PubMed]
  • Bradley A, Evans M, Kaufman MH, Robertson E. Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines. Nature. 1984;309:255–256. doi: 10.1038/309255a0. [PubMed] [Cross Ref]
  • Brandon EP, Idzerda RL, McKnight GS. Targeting the mouse genome: a compendium of knockouts (part I–III) Curr Biol. 1995;5:625–634. doi: 10.1016/S0960-9822(95)00127-8. [PubMed] [Cross Ref]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162:156–159. doi: 10.1016/0003-2697(87)90021-2. [PubMed] [Cross Ref]
  • Clarke AR. Murine genetic models of human disease. Curr Opin Genet Dev. 1994;4:453–460. doi: 10.1016/0959-437X(94)90035-3. [PubMed] [Cross Ref]
  • Cormack BP, Valdivia RH, Falkow S. FACS-optimized mutants of the green fluorescent protein (GFP) Gene. 1996;173:33–38. doi: 10.1016/0378-1119(95)00685-0. [PubMed] [Cross Ref]
  • Dani C, Smith AG, Dessolin S, Leroy P, Staccini L, Villageois P, Darimont C, Ailhaud G. Differentiation of embryonic stem cells into adipocytes in vitro. J Cell Sci. 1997;110:1279–1285. [PubMed]
  • Dinsmore J, Ratliff J, Deason T, Pakzaban P, Jacoby D, Galpern W, Isacson O. Embryonic stem cells differentiated in vitro as a novel source of cells for transplantation. Cell Transplant. 1996;5:131–143. doi: 10.1016/0963-6897(95)02040-3. [PubMed] [Cross Ref]
  • Dinsmore J, Ratcliff J, Jacoby D, Wunderlich M, Lindberg C. Embryonic stem cells as a model for studying regulation of cellular differentiation. Theriogenology. 1998;49:145–151. doi: 10.1016/S0093-691X(97)00409-3. [PubMed] [Cross Ref]
  • Doetschman TC, Eistetter H, Katz M, Schmidt W, Kemler R. The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. J Embryol Exp Morphol. 1985;87:27–45. [PubMed]
  • Drab M, Haller H, Bychkow R, Erdmann B, Lindschau C, Haase H, Morano I, Luft FC, Wobus AM. From totipotent embryonic stem cells to spontaneously contracting vascular smooth muscle cells: a retinoic acid and db-cAMP in vitro differentiation model. FASEB J. 1997;11:905–915. [PubMed]
  • Eistetter HR. Pluripotent embryonal stem cell lines can be established from disaggregated mouse morulae. Dev GrowthDiffer. 1989;31:275–282. doi: 10.1111/j.1440-169X.1989.00275.x. [Cross Ref]
  • Evans MJ, Kaufman MH. Establishment in culture of pluripotential cells from mouse embryos. Nature. 1981;292:154–156. doi: 10.1038/292154a0. [PubMed] [Cross Ref]
  • Fässler R, Meyer M. Consequences of lack of β1 integrin gene expression in mice. Genes Dev. 1995;9:1896–1908. [PubMed]
  • Fässler R, Rohwedel J, Maltsev V, Bloch W, Lentini S, Guan K, Gullberg D, Hescheler J, Addicks K, Wobus AM. Differentiation and integrity of cardiac muscle cells are impaired in the absence of β1 integrin. J Cell Sci. 1996;109:2989–2999. [PubMed]
  • Forrester LM, Nagy A, Sam W, Watt A, Stevenson L, Bernstein A, Joyner AL, Wurst W. An induction of gene trap screen in embryonic stem cells — identification of genes that respond to retinoic acid in vitro. Proc Natl Acad Sci USA. 1996;93:1677–1682. doi: 10.1073/pnas.93.4.1677. [PMC free article] [PubMed] [Cross Ref]
  • Fraichard A, Chassande O, Bilbaut G, Dehay C, Savatier P, Samarut J. In vitro differentiation of embryonic stem cells into glial cells and functional neurons. J Cell Sci. 1995;108:3181–3188. [PubMed]
  • Fürst DO, Osborn M, Nave R, Weber K. The organization of titin filaments in the half-sarcomere revealed by monoclonal antibodies in immunelectron microscopy: a map of ten nonrepetitive epitopes starting at the Z line extends close to the M line. J Cell Biol. 1988;106:1563–1572. doi: 10.1083/jcb.106.5.1563. [PMC free article] [PubMed] [Cross Ref]
  • Gardner RL, Brook FA. Reflections on the biology of embryonic stem (ES) cells. Int J Dev Biol. 1997;41:235–243. [PubMed]
  • Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch. 1981;391:85–100. doi: 10.1007/BF00656997. [PubMed] [Cross Ref]
  • Helgason CD, Sauvageau G, Lawrence HJ, Largman C, Humphries RK. Overexpression of HOXB4 enhances the hematopoietic potential of embryonic stem cells differentiated in vitro. Blood. 1996;87:2740–2749. [PubMed]
  • Hescheler J, Fleischmann BK, Lentini S, Maltsev VA, Rohwedel J, Wobus AM, Addicks K. Embryonic stem cells: a model to study structural and functional properties in cardiomyogenesis. Cardiovasc Res. 1997;36:149–162. doi: 10.1016/S0008-6363(97)00193-4. [PubMed] [Cross Ref]
  • Hogan BLM, Thaller C, Eichele G. Evidence that Hensen's node is a site of retinoic acid synthesis. Nature. 1992;359:237–241. doi: 10.1038/359237a0. [PubMed] [Cross Ref]
  • Hole N and Smith AG (1994) Embryonic stem cells and hematopoiesis. In: Culture of hematopoietic cells, (pp. 235-249) Wiley-Liss, Inc.
  • Hong Y, Winkler C, Schartl M. Pluripotentcy and differentiation of embryonic stem cell lines from the medakafish (Oryzias latipes) Mech Dev. 1996;60:33–44. doi: 10.1016/S0925-4773(96)00596-5. [PubMed] [Cross Ref]
  • Itoh F, Nakane T, Chiba S. Gene expression of MASH-1, MATH-1, neuroD and NSCL-2, basic helix-loop-helix proteins, during neural differentiation in P19 embryonal carcinoma cells. Tohoku J Exp Med. 1997;182:327–336. doi: 10.1620/tjem.182.327. [PubMed] [Cross Ref]
  • Jacob A, Budhiraja S, Reichel RR. Differential induction of HNF-3 transcription factors during neuronal differentiation. Exp Cell Res. 1997;234:277–284. doi: 10.1006/excr.1997.3622. [PubMed] [Cross Ref]
  • Johansson BM, Wiles MW. Evidence for involvement of Activin A and bone morphogenetic protein 4 in mammalian mesoderm and hematopoietic development. Mol Cell Biol. 1995;15:141–151. [PMC free article] [PubMed]
  • Keller GM. In vitro differentiation of embryonic stem cells. Curr Opin Cell Biol. 1995;7:862–869. doi: 10.1016/0955-0674(95)80071-9. [PubMed] [Cross Ref]
  • Klug MG, Soonpa MH, Koh GY, Field LJ. Genetically selected cardiomyocytes from differentiating embryonic stem cells form stable intracardiac grafts. J Clin Invest. 1996;98:216–224. doi: 10.1172/JCI118769. [PMC free article] [PubMed] [Cross Ref]
  • Koh GY, Kim S-J, Klug M, Park K, Soonpa MH, Field L. Targeted expression of transforming growth factor-ß1 in intracardiac grafts promotes vascular endothelial cell DNA synthesis. J Clin Invest. 1995;95:114–121. [PMC free article] [PubMed]
  • Lin Q, Schwarz J, Bucana C, Olson EN. Control of mouse cardiac morphogenesis and myogenesis by transcription factor MEF2C. Science. 1997;276:1404–1407. doi: 10.1126/science.276.5317.1404. [PubMed] [Cross Ref]
  • Lints TJ, Parsons LM, Hartley L, Lyons I, Harvey RP. Nkx-2.5: a novel murine homeobox gene expressed in early heart progenitor cells and their myogenic descendants. Development. 1993;119:419–431. [PubMed]
  • Maltsev VA, Rohwedel J, Hescheler J, Wobus AM. Embryonic stem cells differentiate in vitro into cardiomyocytes representing sinusnodal, atrial and ventricular cell types. Mech Dev. 1993;44:41–50. doi: 10.1016/0925-4773(93)90015-P. [PubMed] [Cross Ref]
  • Maltsev VA, Wobus AM, Rohwedel J, Bader M, Hescheler J. Cardiomyocytes differentiated in vitro from embryonic stem cells developmentally express cardiac-specific genes and ionic currents. Circ Res. 1994;75:233–244. [PubMed]
  • Maltsev VA, Ji GJ, Wobus AM, Fleischmann BK and Hescheler J: Establishment of β-adrenergic modulation of L-type calcium current in the early stages of cardiomyocyte development (submitted) [PubMed]
  • Marshall H, Morrison A, Studer M, Pöpperl H, Krumlauf R. Retinoids and Hox genes. FASEB J. 1996;10:969–978. [PubMed]
  • Martin GR. Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Nat Acad Sci USA. 1981;78:7634–7638. doi: 10.1073/pnas.78.12.7634. [PMC free article] [PubMed] [Cross Ref]
  • Martin GR, Evans MJ. Differentiation of clonal lines of teratocarcinoma cells: formation of embryoid bodies in vitro. Proc Natl Acad Sci USA. 1975;72:1441–1445. doi: 10.1073/pnas.72.4.1441. [PMC free article] [PubMed] [Cross Ref]
  • Miano JM, Cserjesi P, Ligon KL, Periasamy M, Olson EN. Smooth muscle myosin heavy chain exclusively marks the smooth muscle lineage during mouse embryogenesis. Circ Res. 1994;75:803–812. [PubMed]
  • Miller-Hance WC, LaCorbiere M, Fuller SJ, Evans SM, Lyons G, Schmidt C, Robbins J, Chien KR. In vitro chamber specification during embryonic stem cell cardiogenesis. J Biol Chem. 1993;268:25244–25252. [PubMed]
  • Molkentin JD, Lin Q, Duncan SA, Olson EN. Requirement of the transcription factor GATA4 for heart tube formation and ventral morphogenesis. Genes Dev. 1997;11:1061–1072. [PubMed]
  • Monk M. Changes in DNA methylation during mouse embryonic development in relation to X-chromosome activity and imprinting. Philos Trans R Soc Lond B Biol Sci. 1990;326:299–312. [PubMed]
  • Muscat GE, Rea S, Downes M. Identification of a regulatory function for an orphan receptor in muscle: COUP-TF II affects the expression of the myoD gene family during myogenesis. Nucleic Acids Res. 1995;23:1311–1318. [PMC free article] [PubMed]
  • Nagy A, Rossant J, Nagy R, Abramow-Newerly W, Roder JC. Derivation of completely cell culture-derived mice from early passage embryonic stem cells. Proc Natl Acad Sci USA. 1993;90:8424–8428. doi: 10.1073/pnas.90.18.8424. [PMC free article] [PubMed] [Cross Ref]
  • Okabe S, Forsberg-Nilsson K, Spiro AC, Segal M, McKay RD. Development of neuronal precursor cells and functional postmitotic neurons from embryonic stem cells in vitro. Mech Dev. 1996;59:89–102. doi: 10.1016/0925-4773(96)00572-2. [PubMed] [Cross Ref]
  • Pain B, Clark ME, Shen M, Nakazawa H, Sakurai M, Samarut J, Etches RJ. Long-term in vitro culture and characterization of avian embryonic stem cells with multiple morphogenetic potentialities. Development. 1996;122:2339–2348. [PubMed]
  • Palmiter RD, Brinster RL, Hammer RE, Trumbauer ME, Rosenfeld MG, Birnberg NC, Evans RM. Dramatic growth of mice that develop from eggs microinjected with metallothionein-growth hormone fusion genes. Nature. 1982;300:611–615. doi: 10.1038/300611a0. [PubMed] [Cross Ref]
  • Pich U, Pütz D, Wobus AM. Screening method for the measurement of chronotropic effects on cardiac cells. Bioforum. 1997;20:536–540.
  • Potocnik AJ, Kohler H, Eichmann K. Hemato-lymphoid in vivo reconstitution potential of subpopulations derived from in vitro differentiated embryonic stem cells. Proc Natl Acad USA. 1997;94:10295–10300. doi: 10.1073/pnas.94.19.10295. [PMC free article] [PubMed] [Cross Ref]
  • Resnick JL, Bixler LS, Cheng L, Donovan PJ. Long-term proliferation of mouse primordial germ cells in culture. Nature. 1992;359:550–551. doi: 10.1038/359550a0. [PubMed] [Cross Ref]
  • Risau W, Sariola H, Zerwes HG, Sasse J, Ekblom P, Kemler R, Doetschman T. Vasculogenesis and angiogenesis in embryonic stem cell-derived embryoid bodies. Development. 1988;102:471–478. [PubMed]
  • Rohwedel J, Maltsev V, Bober E, Arnold HH, Hescheler J, Wobus AM. Muscle cell differentiation of embryonic stem cells reflects myogenesis in vivo: Developmentally regulated expression of myogenic determination genes and functional expression of ionic currents. Dev Biol. 1994;164:87–101. doi: 10.1006/dbio.1994.1182. [PubMed] [Cross Ref]
  • Rohwedel J, Horak V, Hebrok M, Füchtbauer EM, Wobus AM. M-twist expression inhibits mouse embryonic stem cell-derived myogenic differentiation in vitro. Exp Cell Res. 1995;220:92–100. doi: 10.1006/excr.1995.1295. [PubMed] [Cross Ref]
  • Rohwedel J, Sehlmeyer U, Jin S, Meister A, Wobus AM. Primordial germ cell-derived mouse embryonic germ (EG) cells in vitro resemble undifferentiated stem cells with respect to differentiation capacity and cell cycle distribution. Cell Biol Inter. 1996;20:579–587. doi: 10.1006/cbir.1996.0076. [PubMed] [Cross Ref]
  • Rohwedel J, Kleppisch T, Pich U, Guan K, Jin S, Zuschratter W, Hopf C, Hoch W, Hescheler J, Witzemann V, Wobus AM. Formation of postsynaptic-like membranes during differentiation of embryonic stem cells in vitro. Exp Cell Res. 1998;239:214–225. doi: 10.1006/excr.1997.3903. [PubMed] [Cross Ref]
  • Rohwedel J, Guan K, Zuschratter W, Jin S, Ahnert-Hilger G, Fürst D, Fässler R and Wobus AM (1998b) Loss of β1 integrin function results in a retardation of myogenic, but an acceleration of neuronal differentiation of embryonic stem cells in vitro. Dev Biol 201 (in press). [PubMed]
  • Rose O, Rohwedel J, Reinhardt S, Bachmann M, Cramer M, Rotter M, Wobus AM, Starzinski-Powitz A. Expression of M-cadherin protein in myogenic cells during prenatal mouse development and differentiation of embryonic stem cells in culture. Dev Dyn. 1994;201:245–259. [PubMed]
  • Rudnicki MA, McBurney MW. Cell culture methods and induction of differentiation of embryonal carcinoma cell lines. In: Robertson EJ, editor. Teratocarcinomas and embryonic stem cells — a practical approach. Washington DC, USA: IRL Press Oxford; 1987. pp. 19–49.
  • Rust EM, Westfall MV, Samuelson LC, Metzger JM. Gene transfer into mouse embryonic stem cell-derived cardiac myocytes mediated by recombinant adenovirus. Vitro Cell Dev Biol, Anim. 1997;33:270–276. [PubMed]
  • Sauer H, Hofmann C, Wartenberg M, Wobus AM, Hescheler J. Spontaneous calcium oscillations in embryonic stem cell-derived primitive endodermal cells. Exp Cell Res. 1998;238:13–22. doi: 10.1006/excr.1997.3809. [PubMed] [Cross Ref]
  • Schöler HR, Dressler GR, Balling R, Rohdewohld H, Gruss P. Oct-4: A germline-specific transcription factor mapping to the mouse t-complex. EMBO J. 1990;9:2185–2195. [PMC free article] [PubMed]
  • Schoonjans L, Albright GM, Li JL, Collen D, Moreadith RW. Pluripotential rabbit embryonic stem (ES) cells are capable of forming overt coat color chimeras following injection into blastocysts. Mol Reprod Dev. 1996;45:439–443. doi: 10.1002/(SICI)1098-2795(199612)45:4<439::AID-MRD5>3.0.CO;2-S. [PubMed] [Cross Ref]
  • Solter D, Knowles BB. Monoclonal antibody defining a stage-specific mouse embryonic antigen (SSEA-1) Proc Natl Acad Sci USA. 1978;75:5565–5569. doi: 10.1073/pnas.75.11.5565. [PMC free article] [PubMed] [Cross Ref]
  • Soudais C, Bielinska M, Heikinheimo M, MacArthur CA, Narita N, Saffitz JE, Simon MC, Leiden JM, Wilson DB. Targeted mutagenesis of the transcription factor GATA-4 gene in mouse embryonic stem cells disrupts visceral endoderm differentiation in vitro. Development. 1995;121:3877–3888. [PubMed]
  • Spielmann H, Pohl I, Döring B, Liebsch M and Moldenhauer F (1997) The embryonic stem cell test (EST), an in vitro embryotoxicity test using two permanent mouse cell lines: 3T3 fibroblasts and embryonic stem cells. In: Van Zutphen LFM and Balls M (eds) Animal Alternatives, Welfare and Ethics, (pp. 663-669) Elsevier Science Amsterdam.
  • Srivastava D, Cserjesi P, Olson EN. A subclass of bHLH proteins required for cardiac morphogenesis. Science. 1995;270:1995–1999. [PubMed]
  • Stevens LC. Germ cell origin of testicular and ovarian teratomas. Transplant Proc. 1984;16:502–504. [PubMed]
  • Stewart CL, Gadi I, Bhatt H. Stem cells from primordial germ cells can reenter the germ line. Dev Biol. 1994;161:626–628. doi: 10.1006/dbio.1994.1058. [PubMed] [Cross Ref]
  • Strübing C, Ahnert-Hilger G, Jin S, Wiedenmann B, Hescheler J, Wobus AM. Differentiation of pluripotent embryonic stem cells into the neuronal lineage in vitro gives rise to mature inhibitory and excitatory neurons. Mech Dev. 1995;53:275–287. doi: 10.1016/0925-4773(95)00446-8. [PubMed] [Cross Ref]
  • Thomas KR, Capecchi MR. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell. 1987;51:503–512. doi: 10.1016/0092-8674(87)90646-5. [PubMed] [Cross Ref]
  • Weitzer G, Milner DJ, Kim JU, Bradley A, Capetanaki Y. Cytoskeletal control of myogenesis: a desmin null mutation blocks the myogenic pathway during embryonic stem cell differentiation. Dev Biol. 1995;172:422–439. doi: 10.1006/dbio.1995.8070. [PubMed] [Cross Ref]
  • Wheeler MB. Development and validation of swine embryonic stem cells: a review. Reprod Fertil Dev. 1994;6:563–568. doi: 10.1071/RD9940563. [PubMed] [Cross Ref]
  • Wiles MV, Keller G. Multiple hematopoietic lineages develop from embryonic stem (ES) cells in culture. Development. 1991;111:259–267. [PubMed]
  • Wobus AM, Holzhausen H, Jäkel P, Schöneich J. Characterization of a pluripotent stem cell line derived from a mouse embryo. Exp Cell Res. 1984;152:212–219. doi: 10.1016/0014-4827(84)90246-5. [PubMed] [Cross Ref]
  • Wobus AM, Wallukat G, Hescheler J. Pluripotent mouse embryonic stem cells are able to differentiate into cardiomyocytes expressing chronotropic responses to adrenergic and cholinergic agents and Ca2+ channel blockers. Differentiation. 1991;48:173–182. [PubMed]
  • Wobus AM, Rohwedel J, Maltsev V, Hescheler J. In vitro differentiation of embryonic stem cells into cardiomyocytes or skeletal muscle cells is specifically modulated by retinoic acid. Roux's Arch Dev Biol. 1994;204:36–45. doi: 10.1007/BF00744871. [Cross Ref]
  • Wobus AM, Kleppisch T, Maltsev V, Hescheler J. Cardiomyocyte-like cells differentiated in vitro from embryonic carcinoma cells P19 are characterized by functional expression of adrenoceptors and Ca2+ channels. Vitro Cell Dev Biol, Anim. 1994;30A:425–434. [PubMed]
  • Wobus AM, Rohwedel J, Maltsev V, Hescheler J. Development of cardiomyocytes expressing cardiac-specific genes, action potentials, and ionic channels during embryonic stem cell-derived cardiogenesis. Ann N Y Acad Sci. 1995;752:460–469. [PubMed]
  • Wobus AM, Rohwedel J, Strübing C, Jin S, Adler K, Maltsev V and Hescheler J (1997a) In vitro differentiation of embryonic stem cells. In: Klug S and Thiel R (eds) Methods in Developmental Toxicology and Biology, (pp. 1-17) Blackwell Science Berlin Vienna.
  • Wobus AM, Guan K, Shan J, Wellner MC, Rohwedel J, Ji G, Fleischmann B, Katus HA, Hescheler J, Franz WM. Retinoic acid accelerates embryonic stem cell-derived cardiac differentiation and enhances development of ventricular cardiomyocytes. J Mol Cell Cardiol. 1997;29:1525–1539. doi: 10.1006/jmcc.1997.0433. [PubMed] [Cross Ref]
  • Wobus AM, Guan K. Embryonic stem cell-derived cardiac differentiation: Modulation of differentiation and ‘loss of function’ analysis in vitro. Trends Cardiovasc Med. 1998;8:64–74. doi: 10.1016/S1050-1738(97)00129-1. [PubMed] [Cross Ref]
  • Wobus AM, Guan K and Pich U: In vitro differentiation of embryonic stem cells and analysis of cellular phenotypes, “Gene knockout protocols”, In: Methods in Molecular Biology, Humana Press, Inc. (submitted).
  • Wong H, Anderson WD, Cheng T, Riabowol KT. Monitoring mRNA expression by polymerase chain reaction: the “primer-dropping” method. Anal Biochem. 1994;223:251–258. doi: 10.1006/abio.1994.1581. [PubMed] [Cross Ref]

Articles from Cytotechnology are provided here courtesy of Springer Science+Business Media B.V.


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • PubMed
    PubMed citations for these articles

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...